Compositions and Methods for Treatment of Glaucoma

ABSTRACT

A new use for paracetamol in a method of treating glaucoma, which method comprises administering to a patient in need of such treatment 500 mg to 1000 mg of paracetamol in oral format 4 to 6 hourly. An ophthalmic solution is also provided containing between 0.1 to 5% of paracetamol which is administered as one to two drops in each eye 4 to 6 hourly. The ophthalmic solution further includes one or more of the following excipients: hydroxypropylmethylcellulose, benzalconium chloride, polyacrylic acid such as Teargel®.

FIELD OF THE INVENTION

This invention relates to medicament, more specifically to a medicamentfor use in the treatment of glaucoma.

BACKGROUND TO THE INVENTION

Glaucoma is a well-known eye disorder characterised by abnormally highpressure within the eyeball that leads to loss of retinal ganglion cellsin a characteristic pattern of optic neuropathy. Although raisedintraocular pressure is a significant risk factor for developingglaucoma, there is no set threshold for intraocular pressure that causesglaucoma. Nerve damage may occur at a relatively low pressure in somepatients, while others may have high eye pressure for years and yetnever develop damage. Glaucoma affects one in two hundred people agedfifty and younger, and one in ten over the age of eighty.

If not treated, however, glaucoma leads to permanent damage of the opticnerve and resultant visual field loss, causing impaired vision andsometimes blindness. The loss of visual field often occurs graduallyover a long time and may only be recognized when it is already quiteadvanced.

Several drugs with different mechanisms of action are currentlyavailable for the treatment of glaucoma, each with a specific mechanismof action and efficacy or tolerability. New molecules and mechanisms ofaction are the subject of much research. Cannabis, or marijuana, isknown to reduce the intraocular pressure. However, the widespread use ofcannabis as an illegal recreational substance as well as its sideeffects have limited its application for treating the intraocularpressure in glaucoma.

OBJECT OF THE INVENTION

It is an object of this invention to provide a medicament for use in thetreatment of glaucoma.

SUMMARY OF THE INVENTION

In accordance with this invention there is provided paracetamol for usein a method of treating glaucoma, which method comprises administeringto a patient in need of such treatment an effective amount ofparacetamol (also known as acetaminophen).

According to one aspect of the invention there is provided for theparacetamol to be administered in the form of an ophthalmic solution;for the solution to contain between 0.1 to 5% of paracetamol; and forone to two drops of the solution to be administered to each eye 4 to 6hourly.

According to a second aspect of the invention there is provided for theparacetamol to be administered to a patient in oral format 4 to 6hourly.

The invention further provides an ophthalmic solution including as itsactive ingredient paracetamol.

Further features of the invention provide for the solution to includeparacetamol in the range 0.1 to 5%; and for the solution to furtherinclude one or more of the following excipients:hydroxypropylmethylcellulose, benzalconium chloride, polyacrylic acid;and Teargel®.

The invention also provides for the use of paracetamol in themanufacture of a medicament for use in a method of treating glaucomawhich method comprises administering to a patient in need of suchtreatment an effective amount of paracetamol.

The invention still further provides a method of treating glaucoma whichincludes administering to a patient in need thereof an effective amountof paracetamol.

Further features of the invention provide for the method to include theoral administration of paracetamol, alternately the ophthalmicadministration of paracetamol to a patient.

The invention yet further provides a package which includes at least onedose of paracetamol in an amount effective for the treatment of glaucomaand instructions for the use thereof in a treatment for glaucoma.

Further features of the invention provide for the at least one dose tobe suitable for oral administration, alternately suitable for ophthalmicadministration.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention will be described, by way of example only, with referenceto FIG. 1 which is a two-dimensional line graph showing mean flux valuesof paracetamol in aqueous solution (0.1%) and Teargel® (0.1%) acrosshuman and rabbit corneas where bars represent the standard error of themean.

DETAILED DESCRIPTION WITH REFERENCE TO THE DRAWINGS Treatment ofPatients with Glaucoma

In initial trials, 1000 mg of paracetamol was administered orally 4 to 6hourly to several patients with glaucoma. In all cases the intra-ocularpressure reduction was in the order of 30% after 24 to 48 hours. Thiscompares well with the pressure lowering effect of currently availablecommercial preparations.

Extrapolating from this test, it appears that a dosage in the range of500 to 1000 mg should be sufficient to treat glaucoma. Whilst such dosesevery 4 to 6 hours have been shown to be effective, it is possible thatthis frequency could be reduced.

Although the mechanism of action in the present application is not fullyunderstood it is suspected that it is through paracetamol's indirectactivation of cannabinoid CB1 receptors. The CB1 receptor is expressedin the central nervous system, eye and numerous other tissues and hasbeen recognized as an important therapeutic target for glaucoma. It issuspected that it is through this pathway that paracetamol acts toreduce intra-ocular pressure.

Given the effects of paracetamol on the hepatic and renal systems, anophthalmic solution was proposed as the preferable manner of deliveringan effective amount of paracetamol to the eye in a glaucoma patient.

An ophthalmic solution was made up as follows: 1 mg/ml (0.1%) solutionof paracetamol in phosphate buffered saline (pH=7.4) was used for halfof the experiments and 1 mg/ml (0.1%) solution of paracetamol inphosphate corneas. In more than 2000 corneal penetration experiments ithas been demonstrated that paracetamol does penetrate the cornea well.Corneal penetration is statistically significantly improved when theparacetamol is suspended in polyacrylic acid gel. These tests wereconducted in the following manner.

Materials and Methods Cornea Specimens

Harvested donor comeas, which were deemed unsuitable for transplantationbecause of infections such as hepatitis B and human immunodeficiencyvirus (HIV), were obtained from The Eye Bank Foundation of South AfricaObservatory from 6 patients (mean age±SD, 35±12 years; range, 24-65years). In addition, rabbit corneas were obtained from 6 freshlyslaughtered rabbits (Country Lane Nursery Farm, Kraaifontein, Cape Town,South Africa). Excised corneas were immediately placed inMcCarey-Kaufman solution and transported to a laboratory within 6 hours.The McCarey-Kaufman solution consisted of a stock solution of Medium 199(Sigma Chemical Company, St. Louis, Mo., U.S.A.) to which sodiumbicarbonate, as well as 25 mM of HEPES and 5% dextran, was added beforeusing the solution for the transport of corneal specimens. On arrival inthe laboratory, the corneas were hemisected. One half was usedimmediately, and the other half was snap-frozen in liquid nitrogen andstored at −85° C. for no longer than 6 months before use. At least onepiece of corneal tissue from each specimen was placed in formalin andretained for histologic examination.

No specimens were obtained in which there was clinical evidence of anycorneal disease or pathology that might have influenced the permeabilitycharacteristics of the cornea. The study was approved by the EthicsCommittee of the Faculty of Health Sciences, University of Stellenboschand the Tygerberg Academic Hospital in Cape Town, South Africa.

Permeability Experiments and Histology

Before each permeability experiment, tissue specimens were thawed atroom temperature in phosphate buffered saline (PBS, pH 7.4). Althoughsome damage may have occurred on a cellular level, the processes offreezing, storage, and thawing were demonstrated to have no adverseeffects on the permeability characteristics of corneal tissues in 2previous studies (Van der Bijl et al., 2001; Van der Bijl et al., 2002).Thereafter, corneas were very carefully cut, so as not to damage eitherthe endo- or epithelial surfaces, into 4-mm² sections and mounted inflow-through diffusion cells (exposed areas, 0.039 cm2) as previouslydescribed (Van der Bijl et al., 1997; Van der Bijl et al., 1998; Van derBijl et al., 2000). Each experiment using the paracetamol solution wasrepeated six times for the human and rabbit corneas, respectively.Before commencing each permeability experiment, tissue disks wereequilibrated for 10 minutes with PBS (pH 7.4) at 20° C. in both thedonor and receiver compartments of the diffusion cells.

Following equilibration, the PBS was removed from the donor compartmentand replaced with 1.0 mL of PBS, containing 1 mg/mL (0.1%) paracetamolin PBS at pH 7.4 (w/v) or else 1 mg/mL (0.1%) paracetamol in acommercially available artificial tear gel i.e. Teargel® liquid gel(Restan Laboratories, Bryanston, South Africa). The latter is a highlyviscous clear gel, with an extended tear film break-up time. The gel,after its addition to the donor chamber, was capped completely with atight-fitting teflon disc and 1 mL of PBS deposited on top of the disc.The PBS did not, in any way, mix with the gel, but was simply used toprevent dessication of the gel during the 24-hour experiment. Each gramof the latter gel contains 2 mg of polyacrylic acid and cetrimide 0.01%m/m as a preservative. PBS at 20° C. was pumped through the receivingchambers at a rate of 1.5 ml/h with a ISMATEC® 16 Channel High precisiontubing pump and collected, by means of a ISCO Retriever IV fractioncollector, at 2-h intervals for 24 h. The permeability studies wereperformed under sink conditions, i.e. at the completion of each run theconcentration of paracetamol solution in the acceptor chamber neverreached 10% of that in the donor compartment. Paracetamol containingsamples were collected in appropriate sampling tubes of the fractioncollector. Samples were analyzed at the Division of Pharmacology,Faculty of Health Sciences, Stellenbosch University by HPLC with UVdetection. The collected fractions were analyzed directly aftercompletion of the respective experiment for their paracetamol content.

Calculation of Flux Values

Flux (J) values across membranes were calculated by means of therelationship

J=Q/A×t(ng×cm⁻²×min⁻¹)

where Q indicates quantity of substance crossing membrane (in ng); A,membrane area exposed (in cm²); and t, time of exposure (in minutes).

Steady State Kinetics

When no statistically significant differences (p<0.05; analysis ofvariance and Duncan's multiple range test) between flux values wereobtained over at least two consecutive time intervals, a steady state(equilibrium kinetics) was assumed to have been reached for a particularcorneal specimen.

Statistical Analysis

Non-linear regression analyses (third order polynomials) were performedusing a GraphPad Prism, Version 4, 2003 computer programme. An F testwas used to compare entire curves. A t-test at steady state, was alsoperformed for comparative purposes. A significance level of 5% was usedfor all tests and comparisons.

HPLC Detection of Paracetamol

Permeant-containing effluent samples, collected from the acceptorcompartments of the perfusion apparatus over the 2-hour samplingintervals, were analyzed using an Hewlett Packard 1100 serieshigh-performance binary liquid chromatograph (Agilent Technologies,Waldbronn, Germany) equipped with an Agilent Eclipse (XDB-C18) Zorbaxanalytical column (3.5-μm particle size), 150×4.6 mm (ID). The lattercolumn was preceded by a 30×2.1-mm (ID) 018 guard column (40-μm particlesize). The temperature was maintained at 40° C. and a flow rate of 1.0mL/min was used for the entire run. The mobile phase consisted of amixture of 2 solvents, A (50 mol/L KH₂PO₄, pH 5.4) and B(acetonitrile-isopropanol, 4:1 vol/vol). Three isocratic mixtures of A:Bwere used in sequential order as follows: firstly for 0 to 1.0 min: 90%Solvent A/10% Solvent B; secondly for 1 to 4.0 min: Linear gradient to70% Solvent A/30% Solvent B, thirdly for 4.10 mins: return to use of 90%Solvent A/10% Solvent B. En equilibration time of 2.5 min was used for90% Solvent A/30% Solvent B. All reagents used for the mobile phase wereHPLC grade (Burdick & Jackson, Honeywell International Inc, Muskegon,Mich.), and all inorganic solvents were filtered through a 0.45-μmfilter. Deionized water was used for preparing all aqueous standard andbuffer solutions. Aliquots (15 μL) from each fraction sample wereinjected directly onto the column. Since no extraction procedure wasrequired, it was unnecessary to determine recovery rates. Paracetamolwas detected at 245 nm (retention time, 3.45 mins), while the total runtime was set at 5.0 minutes. Recording and integration of peaks wereperformed by means of an Agilent Chem. Station. Spiked standards overthe expected concentration range of 250 ng/ml to 4000 ng/ml (4.0 μg/ml)were randomly included in each batch on a variable wave Detector(Agilent, Waldbonn, Germany). Calibration was linear over the entireconcentration range (R²=0.9999).

Results Permeability Experiments

Mean flux values for 0.1% paracetamol in aqueous solution (PBS) andTeargel (TG) across the frozen/thawed rabbit and human cornea versustime are shown in FIG. 1. The diffusion rates of paracetamol (in bothPBS and TG) across human comeas were higher in the first few hours ofthe diffusion experiment than the values found across rabbit corneas.Steady state was also reached much earlier for paracetamol diffusionacross human corneas. Teargel as a soluent did not affect the diffusionrate of paracetamol across human corneas as the graphs are nearlyidentical. Teargel does, however, increase the diffusion rate ofparacetamol across rabbit comeas.

Statistically significant differences (P<0.05) were obtained when wholecurves are compared using an F-test (3^(rd) degree polynomial) betweenthe steady-state flux values of: paracetamol (TG) and paracetamol (inPBS) across rabbit corneas (P<0.0001); paracetamol (in PBS) acrossrabbit and human corneas (P<0.0001); and paracetamol (TG) across rabbitand human comeas (P=0.0095). No statistically significant differenceswere found for paracetamol (TG) and paracetamol (in PBS) across humancomeas (P=0.9939).

For paracetamol from 0.1% paracetamol in PBS, steady-state flux valueswere 774±30 ng·cm⁻²·min⁻¹ and 870±76 ng·cm⁻²·min⁻¹ across rabbit andhuman cornea respectively (FIG. 1). For paracetamol from 0.1%paracetamol in TG, steady-state flux values were 860±39 ng·cm⁻²·min⁻¹and 860±33 ng·cm⁻²·min⁻¹ across rabbit and human cornea respectively(FIG. 1).

CONCLUSIONS

As paracetamol has been safely used in humans for more than 100 yearsand its effects on the body are well known, the invention provides apromising breakthrough in the treatment of glaucoma. This isparticularly so for paracetamol in an ophthalmic solution which showsgood corneal permeability of paracetamol. This has the benefits of theparacetamol acting directly on to have a direct effect on thetherapeutic targets for glaucoma as well as reducing its effects on thehepatic and renal systems.

It is envisaged that doses of paracetamol which are effective in thetreatment of glaucoma will be packaged together with instructions forthe use thereof in the treatment of this condition. Such doses could beintended for oral administration and could consist of one or moretablets, capsules or the like. Alternately the doses could be intendedfor ophthalmic administration and could be provided by one or morecontainers of a suitable ophthalmic solution intended for single orrepeated use.

It will be appreciated that many other embodiments of the inventionexist which fall within the scope of the invention, particularly asregards dosage forms and therapeutic regimens.

REFERENCES

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1. Paracetamol for use in a method of treating glaucoma, which methodcomprises administering to a patient in need of such treatment aneffective amount of paracetamol.
 2. Paracetamol according to claim 1wherein the effective amount of paracetamol is between 500 mg and 1000mg.
 3. Paracetamol according to either claim 1 or 2 wherein theparacetamol is administered to the patient in an oral format. 4.Paracetamol according to claim 1 wherein the paracetamol is administeredto the patient in the form of an ophthalmic solution.
 5. Paracetamolaccording to claim 4 wherein the ophthalmic solution contains between0.1 and 5% of paracetamol.
 6. Paracetamol according to either claim 4 or5 wherein the paracetamol is administered as one to two drops of thesolution to each eye 4 to 6 hourly.
 7. An ophthalmic solution includingas its active ingredient paracetamol.
 8. An ophthalmic solutionaccording to claim 7 wherein the paracetamol is present in the range of0.1 to 5%.
 9. The use of paracetamol in the manufacture of a medicamentfor use in a method of treating glaucoma which method comprisesadministering to a patient in need of such treatment an effective amountof paracetamol.
 10. The use according to either claim 9 wherein theparacetamol is administered to the patient in an oral format.
 11. Theuse according to claim 10 wherein the paracetamol is administered to thepatient in the form of an ophthalmic solution.
 12. The use according toclaim 11 wherein the ophthalmic solution contains between 0.1 and 5% ofparacetamol.
 13. The use according to either claim 12 wherein theparacetamol is administered as one to two drops of the solution to eacheye 4 to 6 hourly.
 14. A method of treatment of glaucoma which includesadministering to a patient in need thereof an effective amount ofparacetamol.
 15. A method as claimed in claim 14 wherein the paracetamolis administered orally.
 16. A method as claimed in claim 14 wherein theparacetamol is administered ophthalmically.
 17. A package which includesat least one dose of paracetamol in an amount effective for thetreatment of glaucoma and instructions for the use thereof in atreatment for glaucoma.
 18. A package as claimed in claim 17 wherein theat least one dose of paracetamol is suitable for oral administration.19. A package as claimed in claim 17 wherein the at least one dose ofparacetamol is suitable for ophthalmic administration.